脂肪间充质干细胞（ASC）扩增分化为大量肝细胞并进行肝细胞移植或生物人工肝治疗是补充肝衰竭时肝脏再生能力不足的有效手段，但目前缺乏经济高效的ASC肝向分化方法。MicroRNA（miR）在多种干细胞的分化调控中发挥作用，目前尚无研究全面明确阐明miR在ASC肝向分化过程中的具体机制。课题组前期研究发现，乙肝肝衰竭患者血清可以促进人ASC的肝向分化，体外分化3天即表现出多边形类肝细胞形态；同时通过Illumina Hiseq2500高通量测序发现肝衰竭患者血清中高表达miR-150等8个miR。本项目拟通过瞬时转染技术筛选促进人ASC肝向分化的高表达miR组合；预测miR的靶基因，并采用相关miR过表达、抑制表达和荧光素酶双报告基因系统鉴定其靶基因；利用mRNA高通量测序技术分析经miR诱导分化的ASC中差异表达的信号通路，提出并验证miR促进ASC肝向分化中的具体调控机制。

Adipose-derived mesenchymal stem cells(ASC) differentiation into large amount of hepatocytes for cell transplantation or biological artificial liver is an effective measure to supplement the decreased liver regeneration capacity during liver failure, however currently there is no economic or effective technique to differentiate ASC into hepatocytes. MicroRNAs exert regulatory effect in the differentiation process of multiple stem cells, but the detailed mechanism by which microRNA facilitate hepatic differentiation of ASC has not clarified yet. Our group found that serum of HBV related liver failure patients could facilitate hepatic differentiation of ASC, and when treated 3 days the cells had already displayed polygonal hepatocyte-like morphology. We also found 8 highly expressed microRNAs such as miR-150 in the serum of liver failure patients with Illumina Hiseq2500 high-throughput sequencing technique. In this project, we will use transient transfection techniques to screen for microRNAs that is essential in hepatic differentiation of human ASC; enrich predicted target genes of microRNAs that promote differentiation of ASC by microRNA database search and literature search, and verify the target gene by using microRNA mimic or inhibitor transfection and dual luciferase reporter gene system; analysis the differentially expressed signaling pathways during microRNAs induced ASC differentiation by mRNA high-throughput sequencing to propose and verify specific regulation mechanism of the hepatic differentiation of ASC by microRNA.